From owner-celegans@net.bio.net Sun Nov 22 22:00:00 1998 Path: biosci!biosci!not-for-mail From: Johnny Fares Newsgroups: bionet.celegans Subject: Immunofluorescence Date: 23 Nov 1998 12:13:59 -0800 Organization: Columbia University Lines: 22 Sender: daemon@net.bio.net Approved: wmorgan@acs.wooster.edu Distribution: world Message-ID: <73cfm7$ea4@net.bio.net> NNTP-Posting-Host: net.bio.net I am using the modified Finney-Ruvkun protocol to prepare worms for immunofluorescence. I have no problems with older worms, but I am losing most of L1-L2 worms during the Borate washes. Before Borate (with TTE) the worms spin down to the bottom of the eppendorf tube after one minute spin at 4000rpm. With the borate, I can see most of my worms stuck to the side of the tube even after a more vigourous spin. I tried using slick ep tubes and that didn't help. Any suggestions to help me retain my worms? I suspect that it tis the detergent in the TTE that is keeping the worms from sticking to the sides of the tube. Maybe adding Triton X100 to the Borate buffer, or is that bad chemistry? Hanna (Johnny) Fares 701 West 168 Street HHSC Room 718 Dept. of Biochemistry Columbia University New York, NY, 10032 Tel: 212 305 6931 Fax: 212 305 0383 .